Morphologic And Proteomic Analysis Of Avascular Necrosis Of The Femoral Head Caused By Different Causes

BACKGROUND: Osteonecrosis of the femoral head(ONFH)is a disease caused by different incentives,including the degeneration and necrosis of bone marrow stem cells,osteocyte and bone marrow adipocytes,which is one of the common clinical diseases in department of orthopedics.It is mainly divided into two categories: traumatic osteonecrosis of femoral haead and non-traumatic osteonecrosis of the femoral head.At present,ONFH has become the most common disease of the middle and young.Although a large number of literatures reported that non-traumatic avascular necrosis of femoral head is closely related to many factors,steroid and alcoholic avascular necrosis of the femoral head is generally considered to be the most common cause of avascular necrosis of femoral head.However,the exact etiology and pathogenesis of avascular necrosis of the femoral head are still controversial.The etiology of avascular necrosis of the femoral head is complex,and the ischemic necrosis of the femoral head caused by different causes shows different pathological changes.At present,most studies do not involve the comparison of different causes.Most of the proteomics research on disease in the Department of orthopedics is still in the preliminary study stage.In recent years,the proteomics research on avascular necrosis of the femoral head is mainly focused on the study of serology and bone tissue of experimental animals.The proteomic study of bone tissue in patients with ONFH is rarely reported.OBJECTIVE: The purpose of this study is to investigate the histomorphological changes of bone in ONFH caused by different etiologies.and to detect the trend of bone histomorphology in different regions by micro CT.At the same time,the proteomics analysis of osteonecrosis area and normal bone tissue.Objective to explore the relationship between morphological and proteomic changes of ONFH caused by different etiologies,and the mechanism of ONFH.METHODS: Collection of femoral head specimens from patients with hip arthroplasty which are ONFH by different causes.The specimens of ONFH were cut by the same method,and the morphological analysis of bone tissue and the analysis of proteomic analysis were obtained.In morphological analysis,the bone tissue of the same part was obtained,which included the cartilage of the femoral head,the subchondral bone,the necrotic area,and the bone tissue in the normal area.Micro-CT were used for whole and segmental analysis,so as to get morphological changes of bone tissue and continuous changes of bone tissue morphology in different regions after ONFH.At the same time,the necrotic and normal bone tissues were extracted for proteomic analysis.The difference protein from the necrotic region and the normal area was tested in vitro and in vitro.Bone Mesenchymal Stem Cells(BMSC)was extracted and identified,and the ONFH was established in vitro.The proliferation of bone marrow mesenchymal stem cells was detected by CCK-8.The activity of alkaline phosphatase(ALP),real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the osteogenic differentiation of the cells.Animal models of avascular necrosis of femoral head were established.The effects of avascular necrosis of femoral head on bone histomorphology and proteomics were detected by Micro CT system,fluorescence confocal microscopy,histomorphology and immunohistochemistry.Finally,the relationship between morphologic changes and proteomics of avascular necrosis of the femoral head was discussed.RESULTS:(1)The different morphological changes in avascular necrosis of femoral head caused by different etiological factors may cause the collapse of avascular necrosis of femoral head.(2)There is a large number of bone tissue loss in steroid induced avascular necrosis of the femoral head,and the morphological changes in different pathological sections are smaller.(3)In steroid induced avascular necrosis of the femoral head,the content of COL5A2 in the bone tissue in the necrotic region is significantly decreased.CONCLUSION:(1)The Micro CT analysis showed that the morphological changes of the ONFH were different in different causes.(2)The bone tissue proteins in the normal and necrotic areas of the ONFH were significantly different,mainly on the collagen and bone related proteins.(3)In the steroid induced ONFH,the Collagen type V a2(COL5A2)decreased significantly in the necrotic area compared with the normal bone tissue.